EP3538114A1 - Methods of using human mesenchymal stem cells to effect cellular and humoral immunity - Google Patents

Methods of using human mesenchymal stem cells to effect cellular and humoral immunity

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Publication number
EP3538114A1
EP3538114A1 EP17812104.2A EP17812104A EP3538114A1 EP 3538114 A1 EP3538114 A1 EP 3538114A1 EP 17812104 A EP17812104 A EP 17812104A EP 3538114 A1 EP3538114 A1 EP 3538114A1
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EP
European Patent Office
Prior art keywords
cells
subject
mesenchymal stem
stem cells
population
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP17812104.2A
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German (de)
English (en)
French (fr)
Inventor
Joshua M. Hare
Ana Marie LANDIN
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Longeveron Inc
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Longeveron Inc
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Publication date
Application filed by Longeveron Inc filed Critical Longeveron Inc
Publication of EP3538114A1 publication Critical patent/EP3538114A1/en
Pending legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0662—Stem cells
    • C12N5/0663—Bone marrow mesenchymal stem cells (BM-MSC)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5047—Cells of the immune system
    • G01N33/505—Cells of the immune system involving T-cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5047—Cells of the immune system
    • G01N33/5052—Cells of the immune system involving B-cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/52—Assays involving cytokines
    • G01N2333/525—Tumor necrosis factor [TNF]
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • the present invention relates to methods of administering therapeutically effective amounts of isolated populations of allogeneic mesenchymal stem cells to effect cellular and humoral immunity in subjects suffering from non-ischemic dilated cardiomyopathy.
  • the present invention also relates to methods of administering therapeutically effective amounts of isolated populations of allogeneic mesenchymal stem cells to effect cellular and humoral immunity in subjects suffering from symptoms of aging frailty.
  • Aging frailty poses a very concerning problem for the overall health and well- being of individuals and is characterized as a syndrome of multisystem physiological dysregulation. Aging frailty is a geriatric syndrome characterized by weakness, low physical activity, slowed motor performance, exhaustion, and unintentional weight loss. See Yao, X. et al, Clinics in Geriatric Medicine 27(7 ⁇ :79-87 (2011). Furthermore, there are many studies showing a direct correlation between aging frailty and inflammation. See Hubbard, R E. et al, Biogerontoiogy 11 (5):6i5-(A ⁇ (2010).
  • Immunosenescence is characterized by a low grade, chronic systemic inflammatory state known as inflammaging. See Franceshi, C. et al, Annals of the New York Academy of Sciences 908:244-254 (2000). This heightened inflammatory state or chronic inflammation found in aging and aging frailty leads to immune dy sregulation and a complex remodeling of both innate and adaptive immunity.
  • the T cell and B cell repertoire is skewed resulting in an increase in CD8 + T effector memory cells re- expressing CD45ra (TEMRA) and in the CD19 + late/exhausted memory B cells, and a decrease in the CD8 1 Naive T cells, and in the switched memory B cells (CD27 + ).
  • TEMRA CD45ra
  • Inflammaging has received considerable attention because it proposes a link between immune changes and a number of diseases and conditions (such as aging frailty) common in old age.
  • Circulating inflammatory mediators such as cytokines and acute phase proteins are markers of the low-grade inflammation observed to increase with aging.
  • pro-inflammatory cytokines e.g., TNF- ⁇ , IL-6
  • CSR reduced class switch recombination
  • Immunoglobulin isotype switching is crucial for a proper immune response as the effector functions differ in each isotype.
  • a key player in CSR and somatic hypermutation (SHM) is the enzyme, activation-induced cytidine deaminase (AID), encoded by the Aicda gene.
  • AID's basic function in CSR and SHM is to initiate breaks in the DNA by converting cytosines to uracils in the switch and variable regions of immunoglobulins.
  • E47 encoded by the Tcfe2a (E2A) gene, is a transcription factor belonging to the class I basic helix loop helix (bHLH) proteins, also known as E proteins.
  • E47 Without E47 expression, the B cell specific transcription factors EBF1 (early B cell factor) and Pax-5 (paired box protein) are not expressed. Both E47 and Pax-5 are key transcription factors in early development for the B cell lineage and mature B cell function. See Hagman J. el al., Immunity 27(1):%- ⁇ Q (2007); Horcher M. et al, Immunity 14(6) J79-790 (2001); Riley R.L. et al, Seminars in Immunology 17(5).330-336 (2005).
  • the Pax-5 gene encodes the B cell lineage specific activator protein (BSAP) that is expressed at all stages of B cell differentiation, but not in terminally differentiated B cells.
  • BSAP B cell lineage specific activator protein
  • Pax-5 controls B cell commitment by repressing B lineage inappropriate genes and activating B cell specific genes making Pax-5 the B cell gatekeeper and is exclusively expressed in the B lymphoid lineage from the committed pro-B cell to the mature B cell stage.
  • the B cell specific transcription factor, Pax-5 is not only highly important in early B cell development and B cell lineage commitment, it is also involved in CSR.
  • TNF-a the amount of TNF-a made: (1) depends on the amount of system inflammation and (2) impairs the ability of the same B cells to be stimulated with mitogens or antigens. See Frasca, D. et al., Journal of Immunology 188(1): ⁇ 9-2&6 (2012). Thus, the immune response in subjects suffering from aging frailty is impaired for a number of reasons.
  • NIDCM Non-Ischemic Dilated Cardiomyopathy
  • NIDCM Non-ischemic dilated cardiomyopathy
  • Mesenchymal stem cells are multipotent cells able to migrate to sites of injury, while also being immunoprivileged by not detectably expressing major histocompatibility complex class II (MHC-II) molecules, and expressing MHC-I molecules at low levels.
  • MHC-II major histocompatibility complex class II
  • allogeneic mesenchymal stem cells hold great promise for therapeutic and regenerative medicine, and have been repeatedly shown to have a high safety and efficacy profile in clinical trials for multiple disease processes. See Hare, J.M.
  • mesenchymal stem cells are also a potential source of multiple cell types for use in tissue engineering. See Gong Z. et al. Methods in Mol. Bio. 698:279-294 (2011); Price, A.P. et al, Tissue Engineering Part A 76(8/2581-2591 (2010); and Togel F. etal, Organogenesis 7(2/96-100 (2011).
  • Mesenchymal stem cells have immuno-modulatory capacity. They control inflammation and the cytokine production of lymphocytes and myeloid-derived immune cells without evidence of immunosuppressive toxicity' and are hypo-immunogenic. See Bernardo M.E. etal, Cell Stem Cell 75( ⁇ /392-402 (2013).
  • Mesenchymal stem cells also have the capacity to differentiate not only into cells of mesodermal origin, but into cells of endodermal and ectodermal origin. See Le Blanc K. et al, Exp. Hematol. 37(70/890-896 (2003).
  • mesenchymal stem cells cultured in airway growth media differentiate to express lung-specific epithelial markers, e.g., surfactant protein-C, Clara cell secretory protein, and thyroid transcription factor-1.
  • lung-specific epithelial markers e.g., surfactant protein-C, Clara cell secretory protein, and thyroid transcription factor-1. See Jiang Y. et al. Nature 418(6893)A ⁇ -49 (2002) and Kotton D.N. et al, Development 728(24/5181-5188 (2001).
  • mesenchymal stem cells are reported in the literature to exert a suppressive effect on antibody production as well as proliferation and maturation of B cells. See Uccelli, A. et al. Trends in Immunology 28(5):2 ⁇ 9-226 (2007). Mesenchymal stem cells are also reported to inhibit the generation and function of antigen presenting cells. See Hoogduijn M.J. et al, Int. Immunopharmacology I0(12):1496-1500 (2010). Finally, mesenchymal stem cells are reported to suppress CD4 + and CD8 + T cell proliferation. See Ghannam S. et al, Stem Cell Res. & Ther. 1:2 (2010).
  • the present inventors discovered a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof.
  • the present inventors also discovered a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof.
  • the present inventors identified several biomarkers specific for non-ischemic dilated cardiomyopathy and aging frailty including, but not limited to, the levels of exhausted B cells (CD19 + , CD27 “ , lgD " ), the levels of switched memory B cells (CD19 + , CD27 high , IgD-), the levels of B-cells expressing intracellular TNF- ⁇ , the levels of early activated T-cells (CD3 + , CD69 4 ), the levels of chronic activated T-cells (CD3 + , CD25 + ), the levels of Temra cells (CD45RA + , CCR7 " ), the CD4 + :CD8 + T cell ratio, and the TNF-a concentration in serum.
  • the levels of exhausted B cells CD19 + , CD27 “ , lgD "
  • the levels of switched memory B cells CD19 + , CD27 high , IgD-
  • the levels of B-cells expressing intracellular TNF- ⁇ the levels of early activated
  • One aspect of the invention relates to a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of exhausted B cells (CD19 + , CD27 " , IgD " ) in a sample of the subject' serum decreases by at least 25% as compared to the number of exhausted B cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the non-ischemic dilated cardiomyopathy.
  • Another aspect of the invention relates to a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of switched memory B cells (CD19 + , CD27 high , IgD " ) in a sample of the subject's serum increases by at least 100% as compared to the number of switched memory B cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the nonischemic dilated cardiomyopathy.
  • the number of switched memory B cells CD19 + , CD27 high , IgD "
  • Another aspect of the invention relates to a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of B-cells expressing intracellular TNF-a in a sample of the subject's serum decreases by at least 30% as compared to the number of B-cells expressing intracellular TNF-a in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the nonischemic dilated cardiomyopathy.
  • Another aspect of the invention relates to a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of early activated T-cells (CD3 + , CD69 + ) in a sample of the subject's serum decreases by at least 30% as compared to the number of early activated T- cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the non-ischemic dilated cardiomyopathy.
  • Another aspect of the invention relates to a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of chronic activated T-cells (CD3 + , CD25 + ) in a sample of the subject's serum decreases by at least 70% as compared to the number of chronic activated T-Cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the non-ischemic dilated cardiomyopathy.
  • Another aspect of the invention relates to a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of Temra cells (CD45RA f , CCR7 ' ) in a sample of the subject's serum decreases by at least 40% as compared to the number of Temra cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the non-ischemic dilated cardiomyopathy.
  • Another aspect of the invention relates to a method of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the TNF-a concentration in a sample of the subject's serum decreases by at least 80% as compared to the TNF-a concentration in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the non-ischemic dilated cardiomyopathy.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of exhausted B cells (CD19 + , CD27 " , IgD " ) in a sample of the subject' serum decreases by at least 10% as compared to the number of exhausted B cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of switched memory in a sample of the subject's serum increases by at least 75% as compared to the number of switched memory B cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of B-cells expressing intracellular TNF-a in a sample of the subject's serum decreases by at least 60% as compared to the number of B-cells expressing intracellular TNF-a in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of early activated T-cells (CD3 + , CD69 + ) in a sample of the subject's serum decreases by at least 30% as compared to the number of early activated T-cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of chronic activated T-cells (CD3 + , CD25 + ) in a sample of the subject's serum decreases by at least 75% as compared to the number of chronic activated T-Cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the number of Temra cells (CD45RA + , CCR7 " ) in a sample of the subject's serum decreases by at least 20% as compared to the number of Temra cells in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the TNF-a concentration in a sample of the subject's serum decreases by at least 50% as compared to the TNF-a concentration in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty.
  • Another aspect of the invention relates to a method of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof, wherein the CD4 + :CD8 + T cell ratio in a sample of the subject's serum increases by at least 100% as compared to the CD4 + :CD8 + T cell ratio in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, thereby treating the symptoms of aging frailty
  • the subject is a human. In another embodiment of the invention, the subject is a human who exhibits infiammaging.
  • the mesenchymal stem cells are bone marrow-derived mesenchymal stem cells. In one embodiment of the invention, the mesenchymal stem cells do not express STRO-1. In another embodiment of the invention, the mesenchymal stem cells do not express CD45. In another embodiment of the invention, the mesenchymal stem cells do not express fibroblast surface markers or have a fibroblast morphology. In another embodiment of the invention, the mesenchymal stem cells are not genetically manipulated.
  • the isolated population of allogeneic mesenchymal stem cells is administered in a single dose. In another embodiment of the invention, the isolated population of allogeneic mesenchymal stem cells is administered in multiple doses, e.g., two or more doses. In another embodiment of the invention, the isolated population of allogeneic mesenchymal stem cells is administered at least yearly.
  • the isolated population of allogeneic mesenchymal stem cells is administered systemically. In one embodiment of the invention, the isolated population of allogeneic mesenchymal stem cells is administered by infusion or direct injection. In one embodiment of the invention, the isolated population of allogeneic mesenchymal stem cells is administered intravenously, intraarterially, intramuscularly, intraperitoneally, subcutaneously, intradermally, orally, transendocardially, or intranasally. In a further embodiment, the isolated population of allogeneic mesenchymal stem cells is administered intravenously. In a further embodiment, the isolated population of allogeneic mesenchymal stem cells is administered intramuscularly.
  • the isolated population of allogeneic mesenchymal stem cells is administered at a dose of about 20x10 6 mesenchymal stem cells. In another embodiment of the invention, the isolated population of allogeneic mesenchymal stem cells is administered at a dose of about lOOxlO 6 mesenchymal stem cells. In another embodiment of the invention, the isolated population of allogeneic mesenchymal stem cells is administered at a dose of about 200x10 6 mesenchymal stem cells.
  • the isolated population of allogeneic mesenchymal stem cells are obtained from a human donor and wherein a step of MHC matching of the human donor to the subject is not employed prior to the administration of the isolated population of allogeneic mesenchymal stem cells to the subject.
  • Another aspect of the invention relates to a method of evaluating cellular and humoral immunity status in a subject, comprising:
  • correlating step comprises assigning a likelihood of one or more future changes in immune status to the subject based on the assay result(s); and (4) treating the subject based on the predetermined subpopulation of individuals to which the subject is assigned, wherein the treatment comprises administration of one or more additional doses of an isolated population of allogeneic human mesenchymal stem cells.
  • Another aspect of the invention relates to a method of evaluating cellular and humoral immunity status in a subject, comprising:
  • CD3 + , CD69 + chronic activated T-cells (CD3 + , CD25 + ), Temra cells (CD45RA + , CCR7 " ), the CD4 + :CD8 + T cell ratio, and serum TNF-a by introducing the serum sample obtained from the subject into an assay instrument which (i) contacts the serum sample with one or more antibodies which specifically bind for detection the biomarker(s) which are assayed, and (ii) generates one or more assay results indicating of binding of each biomarker which is assayed to a respective antibody to provide one or more assay results;
  • the correlating step comprises assigning a likelihood of one or more future changes in immune status to the subject based on the assay result(s);
  • the treatment comprises administration of one or more additional doses of an isolated population of allogeneic human mesenchymal stem cells.
  • one or more future changes in immune status comprise one or more of an increase in the number of exhausted B cells (CD 19* CD27 “ , IgD " ), a decrease in the number of switched memory B cells
  • an increase in the number of B-cells expressing intracellular TNF-a an increase in the number of early activated T-cells (CD3 + , CD69*), an increase in the number of chronic activated T-cells (CD3 + , CD25 1 ), an increase in the number of Temra cells (CD45RA , CCR7 " ), a decrease in the CD4 + :CD8 + T cell ratio, and an increase in serum TNF-a.
  • Another aspect of the invention relates to an in vitro method of determining efficacy of treatment of non-ischemic dilated cardiomyopathy in a subject comprising: detennining the levels of one or more biomarkers selected from the group consisting of exhausted B cells (CD19 + , CD27 " , IgD ), switched memory B cells (CD19 + , CD27 high , IgD ), B-cells expressing intracellular TNF-a, early activated T-cells (CD3 + , CD69*), chronic activated T-cells (CD3 + , CD25 + ), Temra cells (CD45RA + , CCR7 " ), and the TNF-a concentration in serum obtained from the subject before and after administration of a population of isolated allogeneic human mesenchymal stem cells to the subject, and comparing the levels of the one or more biomarkers in the serum obtained before and after administration of the population of isolated human mesenchymal stem cells, wherein treatment is efficacious if
  • the number of exhausted B cells decreases by at least 25% as compared to the number of exhausted B cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of switched memory B ( , , g ) increases by at least 100% as compared to the number of switched memory B cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of B-cells expressing intracellular TNF-a decreases by at least 30% as compared to the number of B-cells expressing intracellular TNF-a prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of early activated T-cells decreases by at least 30% as compared to the number of early activated T-cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of chronic activated T-cells decreases by at least 70% as compared to the number of chronic activated T-Cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of Temra cells decreases by at least 40% as compared to the number of Temra cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells, and/or
  • the TNF-a concentration in a sample of the subject's serum decreases by at least 80% as compared to the TNF-a concentration in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells.
  • Another aspect of the invention relates to an in vitro method of determining efficacy of treatment of symptoms of aging frailty in a subject comprising: determining the levels of one or more biomarkers selected from the group consisting of exhausted B cells (CD19 + , CD27 ⁇ IgD ), switched memory B cells (CD19 + , CD27 high , IgD * ), B-cells expressing intracellular TNF-a, early activated T-cells (CD3 + , CD69 + ), chronic activated T-cells (CD3 + , CD25 + ), Temra cells (CD45RA + , CCR7 " ), the CD4 + :CD8 + T cell ratio, and the TNF-a concentration in serum obtained from the subject before and after administration of a population of isolated allogeneic human mesenchymal stem cells to the subject, and comparing the levels of the one or more biomarkers in the serum obtained before and after administration of the population of isolated human mesenchymal stem cells, wherein treatment is
  • the number of exhausted B cells decreases by at least 10% as compared to the number of exhausted B cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of switched memory B cells increases by at least 75% as compared to the number of switched memory B cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of B-cells expressing intracellular TNF-a decreases by at least 60% as compared to the number of B-cells expressing intracellular TNF-a prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of early activated T-cells decreases by at least 30% as compared to the number of early activated T-cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of chronic activated T-cells decreases by at least 75% as compared to the number of chronic activated T-Cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the number of Temra cells decreases by at least 20% as compared to the number of Temra cells prior to administration of the population of isolated allogeneic human mesenchymal stem cells
  • the TNF-a concentration in a sample of the subject's serum decreases by at least 50% as compared to the TNF-a concentration in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells, and/or
  • the CD4+:CD8+ T cell ratio in a sample of the subject's serum increases by at least 100% as compared to the CD4+:CD8+ T cell ratio in a sample of the subject's serum prior to administration of the population of isolated allogeneic human mesenchymal stem cells.
  • Another aspect of the invention relates to the use in vitro of the levels of exhausted B cells (CD19 + , CD27 “ , IgD " ), the levels of switched memory B cells (CD19 + , CD27 hi8h , IgD " ), the levels of B-cells expressing intracellular TNF- ⁇ , the levels of early activated T-cells , the levels of chronic activated T-cells ( the levels of Temra cells and the TNF-a concentration in serum for determining whether treatment for non-ischemic dilated cardiomyopathy is efficacious.
  • Another aspect of the invention relates to the use in vitro of the levels of exhausted B cells the levels of switched memory B cells the ievels of B-cells expressing intracellular TNF- ⁇ , the levels of early activated T-cells the levels of chronic activated T-cells (CD3 + , CD25 1 ), the levels of Temra cells (CD45RA + , CCR7 " ), the CD4 + :CD8 + T cell ratio, and the TNF-a concentration in serum for determining whether treatment for aging frailty is efficacious.
  • FIG. 1 is a Consort Diagram for a randomized, double-blinded, placebo- controlled study investigating the use of allogeneic mesenchymal stem cells for the treatment of older individuals with frailty.
  • FIG. 2 provides baseline patient characteristics for subjects enrolled in the randomized, double-blinded, placebo-controlled study investigating the use of allogeneic mesenchymal stem cells for the treatment of older individuals with frailty.
  • FIG. 3 shows a decrease in TNF-a in older subjects administered allogeneic human mesenchymal stem cells.
  • FIG. 4 shows a decrease in early activated T cells expressing CD69 in older subjects administered allogeneic human mesenchymal stem cells.
  • FIG. 5 shows a decrease in the numbers of chronic/late activated T cells expressing CD2S in older subjects administered allogeneic human mesenchymal stem cells.
  • FIG. 6 shows measurements of CD8+ T cells in older subjects administered allogeneic human mesenchymal stem cells.
  • FIG. 7 shows an increase in the ratio of CD47CD8 1 T cells in older subjects administered allogeneic human mesenchymal stem cells.
  • FIG. 8 shows an increased immunosenescence score in patients with aging frailty who received two infusions of human mesenchymal stem cells.
  • FIG. 9 shows a correlation between baseline serum TNF-a and baseline B cells expressing IC TNF-a .
  • FIG. 10 provides the % of B cells expressing IC TNF-a in Young Control BL, Low TNF:IC TNF BL, and High TNF:IC TNF BL.
  • FIG. 11 provides the % of Switch Memory B cells in Young Control BL, Low TNF:SwMem B cells BL, and High TNF:SwMem B cells BL.
  • FIG. 12 provides the % of Exhausted B cells in Young Control BL, Low TNF:Exh B cells BL, and High TNF:Exh B cells BL.
  • FIG. 13 provides the % of Temra cells in Young Control BL, Low TNFiTemra cells BL, and High TNF:Temra cells BL.
  • FIG. 14 provides the CD4 + /CD8 + T cell ratio in Young Control BL, Low TNF:CD4:CD8 BL, and High TNF:CD4:CD8 BL.
  • FIG. 15 provides the absolute change of serum TNF-a in pg/ml in Placebo Control, Low TNF-a, and High TNF-a.
  • FIG. 16 shows a correlation between baseline serum TNF-a and the change in serum TNF-a.
  • FIG. 17 shows the absolute change of B cells expressing IC TNF-a in Placebo Control, Low TNF-a, and High TNF-a.
  • FIG. 18 shows a correlation between serum TNF-a and change in B cells expressing IC TNF-a.
  • FIG. 19 shows the absolute change of % of Switch Memory B cells in Placebo Control, Low TNF-a, and High TNF-a.
  • FIG. 20 shows the absolute change of % of Exhausted B cells in Placebo Control, Low TNF-a, and High TNF-a.
  • FIG. 21 shows the absolute change of % of Temra T cells in Placebo Control, Low TNF-a, and High TNF-a
  • FIG. 22 shows a correlation between the change in serum TNF-a and the change in Temra T cells.
  • FIG. 23 shows the absolute change in the CD4 + /CD8 + T cell ratio in Placebo Control, Low TNF- ⁇ , and High TNF-a.
  • FIG. 24 shows a correlation between the change in CD4 + /CD8 + T cell ratio and the change in Temra T cells.
  • the present invention is directed to methods of treating non-ischemic dilated cardiomyopathy in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof.
  • the examples demonstrate that in vivo administration of isolated populations of allogeneic human mesenchymal stem cells result in an increase in the percentage of switched memory B cells and a decrease in exhausted B cells in subjects.
  • the examples also demonstrate that in vivo administration of isolated allogeneic human mesenchymal stem cells results in an improvement in the CD4 + :CD8 + T cell ratio in subjects.
  • the levels of B-cells expressing intracellular TNF-a, the levels of early activated T-cells (CD3 + , CD69*), the levels of chronic activated T-cells (CD3 4 , CD25 + ), the levels of Temra cells (CD45RA 4 , CCR7 ), and the TNF-a concentration in serum is reduced in subjects having received infusions of allogeneic human mesenchymal stem cells.
  • the present inventors determined that isolated allogeneic human mesenchymal stem cells favorably altered several immunologic markers typically elevated in chronic inflammation. Restoration of immune competence has clinical relevant in subject who are of higher risk for co-morbid infectious disease.
  • the present invention is directed to methods of treating symptoms of aging frailty in a subject, comprising administering a therapeutically effective amount of a population of isolated allogeneic human mesenchymal stem cells to a subject in need thereof.
  • the examples demonstrate that in vivo administration of isolated populations of allogeneic human mesenchymal stem cells result in an increase in the percentage of switched memory B cells and a decrease in exhausted B cells in subjects.
  • the examples also demonstrate that in vivo administration of isolated allogeneic human mesenchymal stem cells results in an improvement in the CD4 + :CD8 + T cell ratio in subjects.
  • the levels of B-cells expressing intracellular TNF-a, the levels of early activated T- cells (CD3 + , CD69 + ), the levels of chronic activated T-cells (CD3 + , CD25 + ), the levels of Temra cells (CD45RA + , CCR7 " ), and the TNF-a concentration in serum is reduced in subjects having received infusions of allogeneic human mesenchymal stem cells. From these unexpected results, the present inventors determined that isolated allogeneic human mesenchymal stem cells are effective at reducing inflammaging, a prevalent feature in aging frailty.
  • a therapeutically effective amount means an amount that stimulates a B- or T- cell dependent immune response. Such a response is characterized by the ability to elicit significant levels of IgG and opsonic activity.
  • the dosage and number of doses (e.g., single or multiple dose) administered to the subject will vary depending upon a variety of factors, including the route of administration, patient conditions and characteristics (sex, age, body weight, health, size), extent of symptoms, concurrent treatments, frequency of treatment and the effect desired, and the like.
  • the isolated population of allogeneic mesenchymal stem cells is administered as a single dose. In another embodiment, the isolated population of allogeneic mesenchymal stem cells is administered in multiple doses, e.g., two or more doses. In other embodiments, the isolated population of allogeneic mesenchymal stem cells is administered at least yearly.
  • the administration of the isolated population of allogeneic mesenchymal stem cells is repeated, such as at least 1, 2, 3, 4, 5, 6, 8 months after the first administration of the isolated population of allogeneic mesenchymal stem cells, or repeated between 2-4, 2-6, 2-8, months after the first administration of the isolated population of allogeneic mesenchymal stem cells.
  • the isolated population of allogeneic mesenchymal stem cells is administered at a dose of about l
  • the isolated population of allogeneic mesenchymal stem cells is administered at a dose of about 20x10 6 mesenchymal stem cells. In a further embodiment, the isolated population of allogeneic mesenchymal stem cells is administered at a dose of about lOOxlO 6 mesenchymal stem cells. In yet a further embodiment, the isolated population of allogeneic mesenchymal stem cells is administered at a dose of about 200x10 6 mesenchymal stem cells. In further embodiments, the isolated population of allogeneic mesenchymal stem cells is administered
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to increase the ratio of CD4 + :CD8 + T cells in a subject, such as to increase the ratio of CD4 + :CD8 + T cells by at least two-, three, four-, five-, or six-fold as compared to the ratio prior to administration of the isolated population of allogeneic mesenchymal stem cells, i.e., an increase in the ratio of CD4 + :CD8 + T cells by at least 100%, 200%, 300%, 400%, or 500%.
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to increase the number of switched memory B cells (CD19 + , CD27 high , IgD " ) in a subject, such as to increase the number of switched memory B cells by at least two-, three-, four-, or five-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells, i.e., an increase in the number of switched memory B cells (CD19 + , CD27 high ,, IgD " ) by at least 100%, 200%, 300%, or 400%.
  • the number of switched memory B cells as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells increases by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of B cells expressing intracellular TNF-a in a subject, such as to decrease the number by at least two-, three-, four-, five-, or six-fold as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of B cells expressing intracellular TNF-a as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 20%, 30%, 40%, 50%, or 60%.
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of exhausted B cells (CD19 + , CD27 " , IgD " ) in a subject, such as to decrease the number of exhausted B cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of exhausted B cells as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of early activated T-cells (CD3 + , CD69 + ) in a subject, such as to decrease the number of early activated T-cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of early activated T-cells as compared to die number of such T cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 20%, 30%, 40%, or 50%.
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of chronic activated T-cells (CD3 + , CD25 + ) in a subject, such as to decrease the number of chronic activated T-cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of chronic activated T-cells as compared to the number of such T cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of Temra cells (CD45RA + , CCR7 " ) in a subject, such as to decrease the number of Temra cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of Temra cells as compared to the number of such Temra cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 20%, 30%, 40%, or 50%.
  • the therapeutically effective amount of the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the TNF-a concentration in a sample of the subject's serum, such as to decrease the TNF-a concentration by at least two- or three-fold as compared to the TNF-a concentration prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the concentration of TNF-a decreases by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%, as compared to the concentration of TNF-a prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • Other aspects of the invention include methods for evaluating the cellular and humoral immunity status in subjects to determine if the subjects would benefit from the administration of an isolated population of allogeneic mesenchymal stem cells and treating a predetermined subpopulation of subjects based on biomarker data
  • one or more assays could be performed to detect a non-ischemic dilated cardiomyopathy marker selected from the group of exhausted B cells (CD19 + , CD27 " , IgD " ), switched memory B cells (CD19 + , IgD " ), B-cells expressing intracellular TNF-a, early activated T-cells (CD3 4 , CD69 + ), chronic activated T-cells (CD3 f , CD25 4 ), Temra cells (CD45RA 4 , CCR7 ), and serum TNF-a.
  • a non-ischemic dilated cardiomyopathy marker selected from the group of exhausted B cells (CD19 + , CD27 " , IgD " ), switched memory B cells (CD
  • one or more assays could be performed to detect an aging frailty marker selected from the group of exhausted B cells (CD19 + , CD27 “ , IgD " ), switched memory B cells (CD19 + , IgD " ), B-cells expressing intracellular TNF- ⁇ , early activated T-cells (CD3 + , CD69 + ), chronic activated T-cells (CD3 + , CD25 + ), Temra cells (CD45RA + , CCR7 " ), the CD4 + :CD8 + T cell ratio, and serum TNF-a.
  • an aging frailty marker selected from the group of exhausted B cells (CD19 + , CD27 “ , IgD " ), switched memory B cells (CD19 + , IgD " ), B-cells expressing intracellular TNF- ⁇ , early activated T-cells (CD3 + , CD69 + ), chronic activated T-cells (CD3 + , CD25 + ), Temra cells (CD
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to increase the ratio of CD4 + :CD8 + T cells in a subject, such as to increase the ratio of CD4 + :CD8 + T cells by at least two-, three, four-, five-, or six-fold as compared to the ratio prior to administration of the isolated population of allogeneic mesenchymal stem cells, i.e.. an increase in the ratio of CD4 + :CD8 + T cells by at least 100%, 200%, 300%, 400%, or 500%.
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to increase the number of switched memory B cells (CD19 + , CD27 high , IgD " ) in a subject, such as to increase the number of switched memory B cells by at least two-, three-, four-, or five-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells, i.e., an increase in the number of switched memory B cells (CD19 + , C
  • IgD " by at least 100%, 200%, 300%, or 400%.
  • the number of switched memory B cells as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells increases by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of B cells expressing intracellular TNF-a in a subject, such as to decrease the number by at least two-, three-, four-, five-, or six-fold as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of B cells expressing intracellular TNF-a as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 20%, 30%, 40%, 50%, or 60%.
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of exhausted B cells (CD19 + , CD27 " , IgD " ) in a subject, such as to decrease the number of exhausted B cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of exhausted B cells as compared to the number of such B cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%.
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of early activated T-cells (CD3 + , CD69*) in a subject, such as to decrease the number of early activated T-cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of early activated T-cells as compared to the number of such T cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 20%, 30%, 40%, or 50%.
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of chronic activated T-cells (CD3 + , CD25 4 ) in a subject, such as to decrease the number of chronic activated T-cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of chronic activated T-cells as compared to the number of such T cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the number of Temra cells (CD45RA 1 , CCR7 " ) in a subject, such as to decrease the number of Temra cells by at least two- or three-fold as compared to the number prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the number of Temra cells as compared to the number of such Temra cells prior to administration of the isolated population of allogeneic mesenchymal stem cells decreases by at least 10%, 20%, 30%, 40%, or 50%.
  • treatment is demonstrated to be efficacious when the isolated population of allogeneic mesenchymal stem cells is sufficient to decrease the TNF-a concentration in a sample of the subject's serum, such as to decrease the TNF-a concentration by at least two- or three-fold as compared to the TNF-a concentration prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • the concentration of TNF-a decreases by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%, as compared to the concentration of TNF-a prior to administration of the isolated population of allogeneic mesenchymal stem cells.
  • administering a composition may be accomplished by oral administration, injection, infusion, parenteral, intravenous, mucosal, sublingual, intramuscular, intradermal, intranasal, intraperitoneal, intraarterial, subcutaneous absorption or by any method in combination with other known techniques.
  • the isolated population of allogeneic mesenchymal stem cells is administered systemically.
  • the isolated population of allogeneic mesenchymal stem cells is administered by infusion or direct injection.
  • the isolated population of allogeneic mesenchymal stem cells is administered intramuscularly, intravenously, intraarteerially, intraperitoneally, subcutaneously, intradermally, orally, transendocardially, or intranasally.
  • the isolated population of allogeneic mesenchymal stem cells is administered intramuscularly.
  • the isolated population of allogeneic mesenchymal stem cells is administered intravenously.
  • the term "subject" as used herein includes, but is not limited to, humans and non-human vertebrates such as wild, domestic, and farm animals. In some embodiments, the term refers to humans, such as elderly humans >65 years of age, or elderly humans 60-95 years of age. In some embodiments, the human subject exhibits symptoms of aging frailty. In some embodiments, the human subject exhibits inflammaging.
  • allogeneic refers to a cell that is of the same animal species but genetically different in one or more genetic loci as the animal that becomes the "recipient host.” This usually applies to cells transplanted from one animal to another non-identical animal of the same species.
  • the phrase "in need thereof means that the subject has been identified as having a need for the particular method or treatment. In some embodiments, the identification can be by any means of diagnosis. In any of the methods and treatments described herein, the subject can be in need thereof. In some embodiments, the subject is in an environment or will be traveling to an environment in which a particular disease, disorder, or condition is prevalent.
  • Cells are referred to herein as being positive or negative for certain markers.
  • a cell can be negative for CD45, which can also be referred to as CD45 " .
  • the superscript notation " " "” refers to a cell that is negative for the marker linked to the superscript.
  • a marker with the " + " refers to a cell that is positive for that marker.
  • a cell that is referenced as "CD8 + " is positive for CDS.
  • a "+” can also be used to reference the marker as positive.
  • a "-" can also be used to reference the marker as negative.
  • stem cell refers to a cell from the embryo, fetus, or adult that has, under certain conditions, the ability to reproduce itself for long periods or, in the case of adult stem cells, throughout the life of the organism. It also can give rise to specialized cells that make up the tissues and organs of the body.
  • Mesenchymal stem cells are the formative pluripotent blast cells found inter alia in bone marrow, blood, dermis, and periosteum that are capable of differentiating into any kind of the specific types of mesenchymal or connective tissues ⁇ i.e., the tissues of the body that support the specialized elements; particularly adipose, osseous, cartilaginous, elastic, and fibrous connective tissues) depending upon various influences from bioactive factors, such as cytokines.
  • mesenchymal stem cells are isolated from bone marrow of adult humans.
  • the cells are passed through a density gradient to eliminate undesired cell types.
  • the cells can be plated and cultured in appropriate media.
  • the cells are cultured for at least one day or about three to about seven days, and removing non-adherent cells. The adherent cells can then be plated and expanded.
  • Placenta is an excellent readily available source for mesenchymal stem cells.
  • mesenchymal stem cells can be derivable from adipose tissue and bone marrow stromal cells are speculated to be present in other tissues. While there are dramatic qualitative and quantitative differences in the organs from which adult stem cells can be derived, the initial differences between the cells may be relatively superficial and balanced by the similar range of plasticity thej' exhibit.
  • Homogeneous human mesenchymal stem cell compositions which serve as the progenitors for all mesenchymal cell lineages.
  • Mesenchymal stem cells are identified by specific cell surface markers which are identified with unique monoclonal antibodies.
  • the homogeneous mesenchymal stem cell compositions are obtained by positive selection of adherent marrow or periosteal cells which are free of markers associated with either hematopoietic or differentiated mesenchymal cells.
  • These isolated mesenchymal cell populations display epitopic characteristics associated with only mesenchymal stem cells, have the ability to regenerate in culture without differentiating, and have the ability to differentiate into specific mesenchymal lineages when either induced in vitro or placed in vivo at a site of inflammation.
  • pluri potent mesenchymal stem cells are separated from other cells in the bone marrow or other mesenchymal stem cell source.
  • Bone marrow cells may be obtained from iliac crest, femora, tibiae, spine, rib, or other medullary spaces.
  • Other spaces of human mesenchymal stem cells include embryonic yolk sac, placenta, umbilical cord, fetal and adolescent skin, and blood.
  • the human mesenchymal stem cells are identified by the absence of markers.
  • human mesenchymal stem cells useful in the invention include those that are negative for STRO-1 and/or negative for CD45.
  • human mesenchymal stem cells useful in the invention include those that do not express fibroblast surface markers or have a fibroblast morphology.
  • the present invention is directed to a method of enhancing a subject's cellular or humoral immune response, comprising administering to the subject therapeutically effective amounts of an isolated population of human mesenchymal stem cells.
  • the mesenchymal stem cells are not genetically manipulated.
  • the mesenchymal stem cells are obtained from a human donor and wherein a step of MHC matching of the human donor to the subject is not employed prior to the administration of the isolated population of human mesenchymal stem cells.
  • compositions for use in the invention may be formulated using any suitable method.
  • Formulation of cells with standard pharmaceutically acceptable carriers and/or excipients may be carried out using routine methods in the pharmaceutical art. The exact nature of a formulation will depend upon several factors including the cells to be administered and the desired route of administration. Suitable types of formulation are fully described in Remington's Pharmaceutical Sciences, 19 th Edition, Mack Publishing Company, Eastern Pennsylvania, USA.
  • compositions may be prepared together with a physiologically acceptable carrier or diluent.
  • a physiologically acceptable carrier or diluent typically, such compositions are prepared as liquid suspensions of cells.
  • the cells may be mixed with an excipient which is pharmaceutically acceptable and compatible with the active ingredient.
  • excipients are, for example, water, saline, dextrose, glycerol, of the like and combinations thereof
  • the pharmaceutical compositions of the invention may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, and/or adjuvants which enhance effectiveness.
  • the adjuvant comprises human serum albumin (HSA).
  • PlasmaLyte ATM This is a sterile, nonpyrogenic isotonic solution for intravenous administration.
  • Each 100 mL contains 526 mg of Sodium Chloride, USP (NaCl); 502 mg of Sodium Gluconate (CeHnNaO?); 368 mg of Sodium Acetate Trihydrate, USP 37 mg of Potassium Chloride, USP (KG); and 30 mg of Magnesium Chloride, USP It contains no antimicrobial agents.
  • the pH is adjusted with sodium hydroxide. The pH is 7.4 (6.5 to 8.0).
  • the mesenchymal stem cells are not genetically manipulated.
  • the mesenchymal stem cells are cryopreserved.
  • the mesenchymal stem cells can be suspended in cryoprotectant consisting of Hespan® (6% hetastarch in 0.9% sodium chloride) supplemented with 2% HSA and 5% DMSO and then aliquoted into cryopreservation containers for placement in vapor phase nitrogen freezers.
  • the mesenchymal stem cells may be provided in PlasmaLyte ATM supplemented with 1% HSA.
  • Baseline assessments included chemistry and hematology laboratories, echocardiography, and chest, abdominal and pelvic computed tomography scans.
  • hMSCs Human Mesenchymal Stem Cells
  • All allogeneic and autologous human mesenchymal stem cells were manufactured at the University of Miami ISO. See, e.g., Golpanian, S. et al, Physiol. Rev. 96:1127-68 (2016) and Mushtaq, M. et al, J. Cardio. Trans. Res. 7:769-80 (2014). Allogeneic human mesenchymal stem cells were derived from Caucasian male donors mean age 25.4 ⁇ 3.3 years and were between 80 to 90% viable at the time of TESI. The auto- hMSCs were from 11 males with a mean age of 58.0 ⁇ 9.9 and six females with a mean age of 55.0 ⁇ 12.4 years.
  • Injection sites were selected to prioritize safety of the TESI procedure and to distribute sites throughout the accessible myocardial territories. Considerations for site selection included avoidance of the ventricular apex, and optimization of catheter stability prior to needle extension.
  • cPRA results showed that 67% of alio and 92% of auto recipients had no reaction to low cPRA (0-20% cPRA). Twenty-seven % of alio and 8% of auto had a moderate cPRA (21-79% cPRA), and one subject (7%) receiving alio MSCs had a high cPRA response (+80% cPRA).
  • FIG. 1 is a Consort Diagram.
  • FIG. 2 provides baseline patient characteristics.
  • FIGs. 3-7 illustrate the impact of allogeneic human MSCs on immune biomarkers.
  • FIG. 3 shows a decrease TNF-a in subjects administered human MSCs. Both early and late/chronic T cell activation decreases after allogeneic MSC treatment.
  • FIG. 4 shows a decrease in early activated T cells expressing CD69 in subjects administered human MSCs.
  • FIG. 5 shows a decrease in the numbers of chronic/late activated T cells expressing CD25 in subjects administered human MSCs.
  • FIG. 7 shows an increase in the ratio of CD4 + /CD8 + T cells in subjects administered human MSCs.
  • Table 10 are the total Scores for each patient that includes different time-points as well as averages among the patients.
  • the immunosenescence Score improves in subjects with aging frailty after the first injection of human mesenchymal stem cells and continues to stay in an improved state after a second injection of human mesenchymal stem cells.

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AU2019374898A1 (en) * 2018-11-08 2021-06-24 University Of Miami Method of determining responsiveness to cell therapy in dilated cardiomyopathy
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WO2023009537A1 (en) 2021-07-26 2023-02-02 Longeveron Inc. Use of mesenchymal stem cells in treatment of juvenile hypoplastic left heart syndrome
AU2022343620A1 (en) 2021-09-10 2024-01-18 Longeveron Inc. Treatment of aging frailty comprising administering bone marrow derived mesenchymal stem cells

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CN110691600A (zh) 2020-01-14
TWI820753B (zh) 2023-11-01
TW202237819A (zh) 2022-10-01
IL266561B1 (en) 2024-02-01
ZA201903574B (en) 2020-12-23
JP2019534307A (ja) 2019-11-28
JP7276664B2 (ja) 2023-05-18
AU2017358042A1 (en) 2019-05-30
IL266561B2 (en) 2024-06-01
WO2018089752A1 (en) 2018-05-17
JP7491964B2 (ja) 2024-05-28
TW201831681A (zh) 2018-09-01
KR20190084091A (ko) 2019-07-15
CN110691600B (zh) 2024-04-16
JP2022106755A (ja) 2022-07-20

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